Codon optimized expression of HPV 16 E6 renders target cells susceptible to E6-specific CTL recognition

Samorski, Regina; Gissmann, Lutz; Osen, Wolfram

doi:10.1016/j.imlet.2006.07.003

Cited by 23 publications

(15 citation statements)

References 41 publications

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“…All incubation steps were carried out for 1 h at 37°C unless indicated otherwise. For the blocking and dilution of antibodies, 5% skim milk in phosphate-buffered saline (PBS)-0.05% Tween 20 (PBS-T) was used; after each incubation step, plates were washed four times with PBS-T; assays were developed with horseradish peroxidase-conjugated goat anti-mouse antibody (Dianova) and 2,2Ј-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS; Sigma) substrate; the absorbance at 405 nm was measured by using an ELISA reader, the supernatant of a hybridoma clone raised with HPV type 16 E6 as an antigen was used as negative control in triplicate (43); and the cutoff was defined as the absorbance values of the negative controls plus three times the standard deviation.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Identification of B-Cell Epitopes on Virus-Like Particles of Cutaneous Alpha-Human Papillomaviruses

Senger

Becker

Schädlich

et al. 2009

J Virol

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Human papillomavirus (PV) (HPV) types 2, 27, and 57 are closely related and, hence, represent a promising model system to study the correlation of phylogenetic relationship and immunological distinctiveness of PVs. These HPV types cause a large fraction of cutaneous warts occurring in immunocompromised patients. Therefore, they constitute a target for the development of virus-like particle (VLP)-based vaccines. However, the immunogenic structure of HPV type 2, 27, and 57 capsids has not been studied yet. Here we provide, for the first time, a characterization of the B-cell epitopes on VLPs of cutaneous alpha-HPVs using a panel of 94 monoclonal antibodies (MAbs) generated upon immunization with capsids from HPV types 2, 27, and 57. The MAbs generated were characterized regarding their reactivities with glutathione S-transferase-L1 fusion proteins from 18 different PV types, the nature of their recognized epitopes, their isotypes, and their ability to neutralize HPV type 2, 27, 57, or 16. In total, 33 of the 94 MAbs (35%) showed type-specific reactivity. All type-specific MAbs recognize linear epitopes, most of which map to the hypervariable surface loop regions of the L1 amino acid sequence. Four of the generated MAbs neutralized pseudovirions of the inoculated HPV type efficiently. All four MAbs recognized epitopes within the BC loop, which is required and sufficient for their neutralizing activity. Our data highlight the immunological distinctiveness of individual HPV types, even in comparison to their closest relatives, and they provide a basis for the development of VLP-based vaccines against cutaneous alpha-HPVs.Recently licensed prophylactic vaccines confer efficient protection against infections by human papillomavirus (PV) (HPV) types 16 and 18, thereby aiming to prevent approximately 70% of all cervical cancer cases (17, 39). These vaccines are composed of virus-like particles (VLPs), which spontaneously assemble from the major capsid protein L1 via 72 pentamers (capsomeres) as subunits (2,23,26).In the process of vaccine development, monoclonal antibodies (MAbs) proved to be valuable tools for the immunological analysis of recombinantly produced capsids and capsomeres (51) as well as for serological studies (25,49,56). Moreover, the identification and characterization of many neutralizing epitopes of HPV types 11 and 16 have been facilitated by the employment of MAbs (6,11,[30][31][32]41,42,55). Such epitopes to neutralizing antibodies are mostly conformation dependent, but a few neutralizing MAbs that recognize linear epitopes have also been generated (16, 18). Most neutralizing MAbs are HPV type specific due to the hypervariable nature of their respective epitopes, which typically reside in the surface-exposed loop regions of the L1 protein (10). In contrast, crossreactive MAbs targeting rather conserved L1 epitopes are generally nonneutralizing.HPV types 2, 27, and 57 are the three members of Alphapapillomavirus species 4 (20). They are very closely related, and HPV types 2 and 27 hardly fulfill the ...

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Section: Methodsmentioning

confidence: 99%

“…The absorbance at 405 nm was measured by using an ELISA reader. As negative control, the supernatant of a hybridoma clone raised with HPV type 16 E6 as an antigen was used in triplicate (43). The cutoff value was defined as absorbance values of the negative controls plus five times the standard deviation.…”

Section: Methodsmentioning

confidence: 99%

Identification of B-Cell Epitopes on Virus-Like Particles of Cutaneous Alpha-Human Papillomaviruses

Senger

Becker

Schädlich

et al. 2009

J Virol

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“…This strategy can enhance the translation of HPV DNA vaccines in DCs. Recently, it has been demonstrated that the immunization of mice with codon-optimized HPV-16 E6 DNA generated stronger E6-specific immune responses than the vaccination of mice with wild-type E6 DNA [72,73]. …”

Section: Strategies To Enhance the Potency Of Therapeutic Hpv Dna Vacmentioning

confidence: 99%

Therapeutic HPV DNA vaccines

Monie¹,

Tsen

Hung³

et al. 2009

Expert Review of Vaccines

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Human papillomavirus (HPV) has been associated with several human cancers, including cervical cancer, vulvar cancer, vaginal and anal cancer, and a subset of head and neck cancers. The identification of HPV as an etiological factor for HPV-associated malignancies creates the opportunity for the control of these cancers through vaccination. Currently, the preventive HPV vaccine using HPV virus-like particles has been proven to be safe and highly effective. However, this preventive vaccine does not have therapeutic effects, and a significant number of people have established HPV infection and HPV-associated lesions. Therefore, it is necessary to develop therapeutic HPV vaccines to facilitate the control of HPV-associated malignancies and their precursor lesions. Among the various forms of therapeutic HPV vaccines, DNA vaccines have emerged as a potentially promising approach for vaccine development due to their safety profile, ease of preparation and stability. However, since DNA does not have the intrinsic ability to amplify or spread in transfected cells like viral vectors, DNA vaccines can have limited immunogenicity. Therefore, it is important to develop innovative strategies to improve DNA vaccine potency. Since dendritic cells (DCs) are key players in the generation of antigen-specific immune responses, it is important to develop innovative strategies to modify the properties of the DNA-transfected DCs. These strategies include increasing the number of antigen-expressing/antigen-loaded DCs, improving antigen processing and presentation in DCs, and enhancing the interaction between DCs and T cells. Many of the studies on DNA vaccines have been performed on preclinical models. Encouraging results from impressive preclinical studies have led to several clinical trials.

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“…It seems that in cervical cancer cells, most of produced E7 proteins are degraded rapidly by the ubiquitin-proteasome pathway; therefore, they are detectable hardly using common detection protein methods such as immunostaining, immunoprecipitation, or western blot analysis (6). On the other hand, some other studies showed that using optimized codons can improve susceptibility of target cells to E6-drived CTL (cytotoxic T lymphocyte) recognition (7). For induction of HPV-specific T lymphocytes in cervical cancer model, vaccination against determined epitopes is an attractive treatment option.…”

Section: Introductionmentioning

confidence: 99%

Co-Administration of Anti-Angiogenic Peptide and DNA Vaccine in Cervical Cancer Tumor Model

et al. 2017

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Background: Cervical cancer is one of the main causes of women's death in the world. Human papilloma virus (HPV) types 16 and 18 have been known as a cause of more than 2/3 of cervical cancers. New combination treatment strategies based on immune responses against viral antigens are likely to be beneficial. Considering the important role of angiogenesis in nutrition and oxygenation of tumor cells, the inhibition of this process can help tumor clearance.

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Codon optimized expression of HPV 16 E6 renders target cells susceptible to E6-specific CTL recognition

Cited by 23 publications

References 41 publications

Identification of B-Cell Epitopes on Virus-Like Particles of Cutaneous Alpha-Human Papillomaviruses

Identification of B-Cell Epitopes on Virus-Like Particles of Cutaneous Alpha-Human Papillomaviruses

Therapeutic HPV DNA vaccines

Co-Administration of Anti-Angiogenic Peptide and DNA Vaccine in Cervical Cancer Tumor Model

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